## STP FOR CLOMIFENE TABLETS

1.0)  Description: Visual

2.0)  Average Weight: Check weight of 20 tablets at randomly and calculate the average weight by formula :  wt of 20 tablets (gm) x 1000 / 20. found avg. wt in mg

3.0) Uniformity of weight:

Weigh 20 tablets selected at random and calculate the average weight. Not more than two of the individual weights deviate from the average weight by more than the percentage shown in table.

Weigh the tablets individually and calculate the percentage of deviation for each tablet. By using formula = (experimental weight – theoretical weight)    x 100%

Theoretical weight

 Average weight of tablets Percentage deviation More than 80mg but Less than 250mg 7.5% 250mg or More 5%

4.0) Identification Test:

1. When examined in the range 230nm to 360nm, the solution obtained in the assay shows absorption maxima at about 235nm and 292nm.
2. Dissolve a quantity of the powdered tablets containing 5mg of clomifene citrate in 5ml of a mixture of 10 volumes of acetic anhydride and 50 volumes of pyridine and heat in a water-bath; a deep red color is produced.

5.0)      Hardness:

The standard method used for tablet hardness testing is compression testing. The tablet is placed between two jaws that crush the tablet. The machine measures the force applied to the tablet and detects when it fractures. Although compressive force is applied to the tablet, along the diameter of the tablet at right angles to the applied force.

6.0)      Disintegration Time

Unless otherwise stated in the individual monograph, introduce one tablet into each tube and add a disc to each tube. The assembly is suspended in the liquid medium in a suitable vessel, preferably a 1-litre beaker. The volume of liquid is such that the wire mesh at its highest point is at least 15mm below the surface of the liquid, and at its lower point is at least 25mm above the bottom of the beaker. At no time should the top of the basket-rack assembly become submerged. There is a thermostatic arrangement for heating the liquid and maintaining the temperature at 37±2°.

If 1 or 2 tablets fail to disintegrate, repeat the test on 12 additional tablets; not less than 16 of the total of 18 tablets tested disintegrate.

If the tablets adhere to the disc and the preparation under examination fails to comply, repeat the test omitting the disc. The preparation complies with the test if all the tablets in the repeat test disintegrate.

7.0)      Dimension of Tablet:

Length, breadth and thickness are determined by vernier in mm

8.0)      Friability:

The test is applicable to compressed tablets and is intended to determine the physical strength of tablets. Tablets with a unit weight equal to or less than 650 mg, take a sample of whole tablets corresponding as near as possible to 6.5g. For tablets with a unit weight of more than 650mg, take a sample of 10 whole tablets. The tablets should be carefully dedusted prior to testing. Accurately weigh the tablet sample, and place the tablets in the drum. Rotate the drum 100 times and remove the tablets. Remove any loose dust from the tablets as before, and accurately weigh.

The test is run only once unless the results are difficult to interpret or if the weight loss is greater than the target value, in which case, the test is repeated twice and the mean of the three tests is determined.

Formula: initial wt. – after friability wt. x100 / initial wt.

A maximum loss of weight (From a single test or from the mean of three tests) not greater than 1.0 percent is acceptable for most tablets.

9.0)      Dissolution:

Apparatus: Basket

Medium: 900ml of water,

Time: 45 minutes

Speed: 100 rpm

Limit: NLT 75% of the stated amount of Clomifene citrate..

Withdraw a suitable volume of medium and filter promptly through a membrane filter disc having an average pore diameter not greater than 1.0µm, rejecting the first 1ml of the filtrate. Dilute a suitable volume of the filtrate with 0.1 M HCl. Measure the absorbance of the resulting solution at the maximum at about 232nm. Calculate the content of clomifene citrate in the medium taking 317 as the specific absorbance at 232nm.

10.0)      Leak test:

The apparatus is used to test for the integrity of packed strips, blisters and Alu-Alu Blister pack containing tablets. Ensure apparatus bath is filled with purified water up to mark indicated and add 0.5% crystal violet solution in water. Samples are placed into the desiccators and the lid is placed in position. The pump starts to produce a vacuum 15inHg inside the desiccators and the vacuum is held for 1 minute. The sample remains at the required vacuum level for given time interval buzzer will sound after time is over and will cut off the vacuum pump. As the package is immersed in a colored dye solution the venting of the desiccators will allow any holes to be penetrated by the dye and the contents of the flexible packaging will also be stained with the same coloring material.

Examine all the strips for any leakage by opening the pockets manually. If anyone pocket shows evidence of leakage, reject the sample, stop the Blister / Strip machine and immediately take corrective action.

10.0)    Z-isomer:

30 to 50 per cent of the content of clomifene citrate as determined in the assay.

Determine by liquid chromatography.

Test solution: Shake a quantity of the powdered tablets containing about 50mg of clomifene citrate with 50ml of 0.1 M HCl for 10 minutes and filter. To 25ml of the filtrate add 5ml of a 1 M sodium hydroxide and extract with 3 quantities, each of 25ml, of ethanol-free chloroform. Wash the combined extracts with 10ml of water, dry over anhydrous sodium sulphate and add sufficient ethanol-free chloroform to produce 100ml. To 20ml of the resulting solution add 0.1 ml of triethylamine and sufficient hexane to produce 100ml.

Chromatographic system:

• A stainless steel column 30cm X4mm, packed with porous silica particles (10µm) (such as parasol).
• Mobile phase: a mixture of ethanol-free chloroform and hexane, each containing 0.10 per cent v/v of triethylamine, adjusted so that the baseline separation is obtained between E- and Z-isomers of clomifene (a mixture of 20 volumes of ethanol-free chloroform and 80 volumes of hexane is suitable),
• Flow rate: 2ml per minute,
• Spectrophotometer set at 302nm,
• Injection volume: about 50µl

Stabilise the system by passing about 250ml of the mobile phase.

Inject the test solution. In the chromatogram a peak due to the E-isomer precedes that due to the Z-isomer of clomifene. The test is not valid unless baseline separation is achieved between E- and Z-clomifene and the column efficiency is greater than 10000 theoretical plates per metre determine using the peak due to E-isomer.

Calculate the percentage of Z-isomer from the expression 100Az/(1.08 AE+Az) where Az and AE are the area of the peaks due to the Z-and E-isomers respectively.

11.0)    Assay: Determined by UV spectroscopy.

Weigh and powder 20 tablets. Weigh accurately a quantity of the powder containing about 50mg of clomifene acid prepared in a 30 per cent v/v solution of 2-propanol ( instead of water normally used for the purpose as solvent). Dilute to 100ml with the propanolic HCl and filter. Dissolve 5.0ml of the filtrate to 100.0ml with 0.1 M HCl and measure the absorbance of the resulting solution at the maximum at about 292nm, using a solution prepared by diluting 5ml of the propanolic HCl to 100ml with 0.1 M HCl as the blank. Calculate the content of Clomifene citrate taking 175 as the specific absorbance at 292nm.

Alternative Method: By UV Spectrophotometer

Test Solution: Weigh and powder 20 tablets. Weight sample equivalent to 25mg of Clomifene citrate 100ml volumetric flask add 20ml methanol into it and sonicate to dissolve the test sample. Make up to mark with methanol. Filter and dilute 2ml of this solution to 25ml in 0.1 M HCl.

Reference Solution: Weigh 25mg of reference standard of Clomifene in 100ml of volumetric flask. Add 20ml methanol into it and sonicate to dissolve. Make up to the mark with methanol. Dilute 2ml of this solution to 25ml with 0.1 M HCl.

Check Absorbance of test as well as reference standard at about 292nm in UV Spectrophotometer.

Calculate the content of Clomifene citrate in the tablets.

Formula:

Abs. of Test      STD Wt.(mg)  2            100               25       Potency

—————–X————–X——-X—————X——–X———–X Average wt.

Abs. of STD        100              25      Test Wt.(mg)     2           100

Acceptance criteria: 92.5%-107.5%

13.0)    MICROBIOLOGICAL PURITY

Perform the test according to requirements of IP,

Total aerobic Microbial count (TAMC): NMT 103 CFU/g

Total combined yeasts/Moulds count (TYMC): NMT 102 CFU/g

Pathogens: in 1gm drug.

Escherichia Coli – Should be absent

Pseudomonas aeroginosa – Should be absent

Salmonella – Should be absent

Staphylococcus aureus– Should be absent

Abbreviations:

Wt.: Weight

mg: Miligram

ml: Milileter

STD: Standard

inHg: Inch of Mercury

rpm: Rounds per minute

CFU: Colony forming unit

HCl: Hydrochloric acid

w/v: Weight/Volume

Abs. Absorbance